RSS-Feed abonnieren
DOI: 10.1055/s-0038-1656377
Mass Contents of Inositol 1,4,5-Trisphosphate and 1,2-Diacylglycerol in Human Platelets Stimulated with a Thromboxane Analogue and Thrombin
Publikationsverlauf
Received 13. Dezember 1991
Accepted after revision 09. April 1992
Publikationsdatum:
04. Juli 2018 (online)
![](https://www.thieme-connect.de/media/10.1055-s-00035024/199203/lookinside/thumbnails/10-1055-s-0038-1656377-1.jpg)
Summary
Mass contents of inositol 1,4,5-trisphosphate (IP3) and 1,2-diacylglycerol (DG) were measured in U46619-stimulated human platelets. 1 µM of U46619 induced maximum responses in aggregation, 5-hydroxytryptamine (5HT) secretion and increase in intracellular free Ca2+ concentration ([Ca2+]i). Aggregation was almost comparable to that induced by maximal dose (1 U/ml) of thrombin, while 5HT release was almost half. The initial [Ca2+]i peak in response to U46619 was about half of thrombin stimulation. Production of IP3 and DG was, however, less than one tenth of that seen in thrombin stimulation. The profile (time course and concentration-dependency) of IP3 formation did not correlate with that of [Ca2+]i, suggesting that U46619 stimulates IPs-dependent and -independent Ca2+ mobilization. DG production was small but sustained for more than 5 min. These findings support the recent hypothesis that aggregation is regulated by a delayed accumulation of DG. The low level of 5HT secretion could be explained by the low production of second messengers, IP3 and DG.
-
References
- 1 Armstrong RA, Jones RL, Wilson NH. Ligand binding to thromboxane receptors on human platelets: correlation with biological activity. Br J Pharmacol 1983; 79: 953-964
- 2 Morinelli TA, Niewiarowski S, Daniel JL, Smith JB. Receptor-mediated effects of a PGH2 analogue (U46619) on human platelets. Am J Physiol 1987; 253: H1035-H1043
- 3 Pollock WK, Armstrong RA, Brydon LJ, Jones RL, MacIntyre DE. Thromboxane-induced phosphatidate formation in human platelets. Relationship to receptor occupancy and to changes in cytosolic free calcium. Biochem J 1984; 219: 833-842
- 4 Siess W, Boehlig B, Weber PC, Lapetina EG. Prostaglandin endoperoxide analogues stimulate phospholipase C and protein phosphorylation during platelet shape change. Blood 1985; 65: 1141-1148
- 5 Brass LF, Shaller CC, Belmonte EJ. Inositol 1,4,5-trisphosphate-induced granule secretion in platelets. Evidence that the activation of phospholipase C mediated by platelet thromboxane receptors involves a guanine nucleotide binding protein-dependent mechanism distinct from that of thrombin. J Clin Invest 1987; 79: 1269-1275
- 6 Kawahara Y, Fukuzaki H, Kaibuchi K, Tsuda T, Hoshijima M, Takai Y. Activation of protein kinase C by the action of 9,11-epithio-11,12-methano-thromboxane A2 (STA2), a stable analogue of thromboxane A2, in human platelets. Thromb Res 1986; 41: 811-818
- 7 Palmer S, Hughes KT, Lee DY, Wakelam MJO. Development of a novel, Ins(1,4,5)P3-specific binding assay, Its use to determine the intracellular concentration of Ins(1,4,5)P3 in unstimulated and vasopressin-stimulated rat hepatocytes. Cell Signalling 1989; 1: 147-156
- 8 Preiss J, Loomis CR, Bishop WR, Stein R, Niedel JE, Bell RM. Quantitative measurement of sn-1,2-diacylglycerols present in platelets, hepatocytes, and ras- and sis-transformed normal rat kidney cells. J Biol Chem 1986; 261: 8597-8600
- 9 Nakashima S, Suganuma A, Matsui A, Nozawa Y. Thrombin induces a biphasic 1,2-diacylglycerol production in human platelets. Biochem J 1991; 275: 355-361
- 10 Born GVR. Aggregation of blood platelets by adenosine diphosphate and its reversal. Nature (London) 1962; 194: 927-929
- 11 Nakashima S, Tohmatsu T, Hattori H, Okano Y, Nozawa Y. Inhibitory action of cyclic GMP on secretion, polyphosphoinositide hydrolysis and calcium mobilization in thrombin-stimulated human platelets. Biochem Biophys Res Commun 1986; 135: 1099-1104
- 12 Sage SO, Merritt JE, Hallam TJ, Rink TJ. Receptor-mediated calcium entry in fura-2-loaded human platelets stimulated with ADP and thrombin. Dual-wavelengths studies with Mn2+ . Biochem J 1989; 258: 923-926
- 13 Bligh EG, Dyer WJ. A rapid method of total lipid extraction and purification. Can J Biochem Physiol 1959; 37: 911-917
- 14 Zschauer A, van Breemen C, Bühler FR, Nelson MT. Calcium channels in thrombin-activated human platelet membrane. Nature (London) 1988; 334: 703-705
- 15 Kaibuchi K, Tsuda T, Kikuchi A, Tanimoto T, Takai Y. Enhancement of collagen-induced phosphoinositide turnover by thromboxane A2 analogue through Ca2+ mobilization in human platelets. FEBS Lett 1985; 192: 104-108
- 16 Brass LF, Joseph SK. A role for inositol triphosphate in intracellular Ca2+ mobilization and granule secretion in platelets. J Biol Chem 1985; 260: 15172-15179
- 17 Daniel JL, Dangelmaier CA, Smith JB. Formation and metabolism of inositol 1,4,5-trisphosphate in human platelets. Biochem J 1987; 246: 109-114
- 18 Traver AP, King WG, Rittenhouse SE. Inositol 1,4,5-trisphosphate and inositol 1,2-cyclic 4,5-trisphosphate are minor components of total mass of inositol trisphosphate in thrombin-stimulated platelets. Rapid formation of inositol 1,3,4-trisphosphate. J Biol Chem 1987; 262: 17268-17271
- 19 Houslay MD, Bojanic D, Wilson A. Platelet activating factor and U44069 stimulate a GTPase activity in human platelets which is distinct from the guanine nucleotide regulatory proteins, Ns and Ni. Biochem J 1986; 234: 737-740
- 20 Shenker A, Goldsmith P, Unson CG, Spiegel AM. The G protein coupled to the thromboxane A2 receptor in human platelets is a member of the novel Gq family. J Biol Chem 1991; 266: 9309-9313
- 21 Taylor SJ, Chae HZ, Rhee SG, Exton JH. Activation of the β1 isozyme of phospholipase C by α subunits of the Gq class of G proteins. Nature (London) 1991; 350: 516-518
- 22 Takahara K, Murray R, FitzGerald GA, Fitzgerald DJ. The response to thromboxane A2 analogues in human platelets. Discrimination of two binding sites linked to distinct effector systems. J Biol Chem 1990; 265: 6836-6844
- 23 Sage SO, Rink TJ. The kinetics of changes in intracellular calcium concentration in fura-2-loaded human platelets. J Biol Chem 1987; 262: 16364-16369
- 24 Jones GD, Gear ARL. Subsecond calcium dynamics in ADP- and thrombin-stimulated platelets: A continuous-flow approach using Indo-1. Blood 1988; 71: 1539-1543
- 25 Authi KS, Evenden BJ, Crawford N. Metabolic and functional consequences of introducing inositol 1,4,5-trisphosphate into saponin-permeabilized human platelets. Biochem J 1986; 233: 707-718
- 26 Werner MH, Hannun YA. Delayed accumulation of diacylglycerol in platelets as a mechanism for regulation of onset of aggregation and secretion. Blood 1991; 78: 435-444